TY  - JOUR
A1  - Wieferig, Jan-Philip
A1  - Mills, Deryck J.
A1  - Kühlbrandt, Werner
T1  - Devitrification reduces beam-induced movement in cryo-EM
T2  - IUCrJ
N2  - As cryo-EM approaches the physical resolution limits imposed by electron optics and radiation damage, it becomes increasingly urgent to address the issues that impede high-resolution structure determination of biological specimens. One of the persistent problems has been beam-induced movement, which occurs when the specimen is irradiated with high-energy electrons. Beam-induced movement results in image blurring and loss of high-resolution information. It is particularly severe for biological samples in unsupported thin films of vitreous water. By controlled devitrification of conventionally plunge-frozen samples, the suspended film of vitrified water was converted into cubic ice, a polycrystalline, mechanically stable solid. It is shown that compared with vitrified samples, devitrification reduces beam-induced movement in the first 5 e Å−2 of an exposure by a factor of ∼4, substantially enhancing the contribution of the initial, minimally damaged frames to a structure. A 3D apoferritin map reconstructed from the first frames of 20 000 particle images of devitrified samples resolved undamaged side chains. Devitrification of frozen-hydrated specimens helps to overcome beam-induced specimen motion in single-particle cryo-EM, as a further step towards realizing the full potential of cryo-EM for high-resolution structure determination.
KW  - cryo-EM
KW  - devitrification
KW  - beam-induced movement
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/75789
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-757891
SN  - 2052-2525
VL  - 8
IS  - 2
SP  - 186
EP  - 194
PB  - International Union of Crystallography
CY  - Chester
ER  -